JP7475395B2 - Compositions and methods of using nintedanib to treat ocular diseases involving abnormal neovascularization - Google Patents

Description

CLAIM OF PRIORITY This application claims the benefit of U.S. Provisional Patent Application No. 62/344,878, filed June 2, 2016, and U.S. Provisional Patent Application No. 62/344,870, filed June 2, 2016, the entire contents of each of which are incorporated herein by reference.

TECHNICAL FIELD The present disclosure relates to ophthalmic compositions and methods using nintedanib for the treatment and prevention of graft rejection in high-risk corneal transplant patients and for the treatment of ophthalmic diseases involving abnormal neovascularization in the anterior segment of the eye.

2. Background Abnormal neovascularization is involved in many diseases in the anterior segment of the eye. Abnormal neovascularization is involved in graft rejection in high-risk corneal transplant patients. Current treatments for many of these indications are in need of improvement. The methods disclosed herein address the problems of current treatments and provide improved treatments for these diseases.

In certain aspects, the present disclosure provides a method for treating an ocular disease associated with abnormal neovascularization in the anterior segment of the eye, comprising administering an effective amount of nintedanib or a pharma- ceutically acceptable salt thereof to an eye of a subject in need of such treatment. In certain aspects, the disclosed method treats, prevents, or delays the onset of graft rejection in corneal transplant patients. For example, the disclosed method treats, prevents, or delays the onset of graft rejection in corneal transplant patients at high risk of graft rejection. In certain aspects, the disclosed method is performed pre-operatively, simultaneously with, or post-operatively to prevent graft rejection in high-risk corneal transplants.

In certain aspects, nintedanib is administered in the form of a topical ophthalmic formulation that is administered locally to the affected eye. In certain aspects, the concentration of nintedanib in the formulation is 0.001% to 10% of the total weight or volume of the composition. For example, an aqueous composition contains 0.001%, 0.01%, 0.1%, 0.5%, 1.0%, 1.5%, 2.0%, 5.0%, or up to 10% nintedanib. In certain aspects, the topical ophthalmic formulation is a solution, suspension, gel, or emulsion. In another aspect, nintedanib is administered in the form of an implant or semi-solid sustained release formulation that is inserted into the affected eye. In certain aspects, the amount of nintedanib in the implant is 1 μg to 100 mg.

The term "subject" refers to an animal or human, or one or more cells derived from an animal or human. Preferably, the subject is a human. Subjects may also include non-human primates. A human subject may be known as a patient.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials used in the present invention are described herein; however, other suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and are not intended to be limiting. All publications, patent applications, patients, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[The present invention 1001]
A method for treating ophthalmic indications associated with abnormal neovascularization comprising administering to the eye of a subject an effective amount of nintedanib, or a pharma- ceutically acceptable salt thereof, in the form of a topical eye drop or in the form of an implant.
[The present invention 1002]
1001. The method of claim 1001, wherein the indication involving abnormal neovascularization is corneal graft rejection after a high-risk patient has undergone a corneal transplant.
[The present invention 1003]
The method of claim 1001, wherein nintedanib is administered in an amount effective to prevent graft rejection by inhibiting abnormal neovascularization and inflammation in the eye of the subject.
[The present invention 1004]
1001. The method of claim 1001, wherein said indication involving abnormal neovascularization is graft-versus-host disease.
[The present invention 1005]
1001. The method of claim 1001, wherein the indication involving abnormal neovascularization is atopic conjunctivitis.
[The present invention 1006]
The method of claim 1001, wherein the indication involving abnormal neovascularization is ocular rosacea.
[The present invention 1007]
The method of claim 1001, wherein the indication involving abnormal neovascularization is ocular pemphigoid.
[The present invention 1008]
1001. The method of claim 1001, wherein said indication involving abnormal neovascularization is Lyell's syndrome.
[The present invention 1009]
1001. The method of claim 1001, wherein said indication involving abnormal neovascularization is neovascularization induced by a viral infection, a bacterial infection, a fungal infection, or a parasitic infection.
[The present invention 1010]
The method of claim 1001, wherein the indication involving abnormal neovascularization is contact lens-induced neovascularization.
[The present invention 1011]
The method of claim 1001, wherein the indication involving abnormal neovascularization is an ulcer.
[The present invention 1012]
1001. The method of claim 1001, wherein the indication involving abnormal neovascularization is alkali burn.
[The present invention 1013]
1001. The method of claim 1001, wherein said indication involving abnormal neovascularization is stem cell deficiency.
[The present invention 1014]
The method of the present invention 1001, wherein the indication involving abnormal neovascularization is pinguecula.
[The present invention 1015]
The method of claim 1001, wherein the indication involving abnormal neovascularization is neovascular glaucoma.
[The present invention 1016]
The method of claim 1001, wherein the indication involving abnormal neovascularization is dry eye disease.
[The present invention 1017]
1001. The method of claim 1001, wherein said indication involving abnormal neovascularization is Sjogren's syndrome.
[The present invention 1018]
The method of the present invention 1001, wherein the indication involving abnormal neovascularization is meibomian gland dysfunction.
[The present invention 1019]
The method of the present invention 1001, wherein the indication involving abnormal neovascularization is Stevens-Johnson syndrome.
[The present invention 1020]
The method of claim 1001, wherein the indication involving abnormal neovascularization is a tumor in the eye.
[The present invention 1021]
The method of claim 10, wherein nintedanib is administered in the form of a topical ophthalmic formulation or in the form of an ocular implant.
[The present invention 1022]
The method of claim 1021, wherein said ocular implant is in the form of a semi-solid or solid sustained release implant.
[The present invention 1023]
The method of claim 1022, wherein the implant is inserted into the eye of the subject.
[The present invention 1024]
The method of claim 1021, wherein said topical ophthalmic preparation is a topical eye drop.
[The present invention 1025]
The method of claim 1021, wherein the topical ophthalmic formulation is a solution, suspension, cream, ointment, gel, gel-forming liquid, suspension containing liposomes or micelles, a spray formulation, or an emulsion.
Other features and advantages of the invention will become apparent from the following detailed description and drawings, and from the claims.

FIG. 1 is a schematic diagram illustrating an exemplary mechanism for preventing graft rejection in high-risk corneal transplant patients according to the present disclosure. Figures 2A and 2B are graphs showing that corneal neovascularization was reduced in the presence of nintedanib in a rabbit corneal suture model. Figure 2A provides the results on day 12, and Figure 2B provides the results on day 14. The area of corneal neovascularization in each treatment group is shown (CBT-1 = nintedanib ophthalmic formulation: 0.2% CBT-1 BID, 0.2% CBT-1 TID; 0.05% CBT-1 BID, 0.0.5% CBT-1 TID; vehicle control TID). T-test significance levels comparing each group to vehicle are indicated by asterisks.

DETAILED DESCRIPTION Corneal transplantation is a common surgical procedure. Although the overall success rate of corneal transplantation is good, graft failure remains a problem in some high-risk patients. These patients have strong inflammation and neovascularization at the host transplant site, which enhances the immune response and rejection of the allograft (Yu et al. World J Transplant. 2016; 6(1): 10-27). Oral immunosuppressants are sometimes used to reduce the risk of graft failure, but they have systemic side effects. The disclosed method prevents graft rejection in high-risk patients by inhibiting excessive neovascularization mediated by vascular endothelial growth factor ("VEGF") and platelet-derived growth factor ("PDGFR"), and attenuating the immune response associated with VEGF and fibroblast growth factor ("FGF"). The mechanism is illustrated in Figure 1.

In addition to corneal graft rejection, the disclosed methods can be used to treat any ocular indication involving abnormal neovascularization in the anterior segment of the eye. These indications include graft-versus-host disease, atopic conjunctivitis, ocular rosacea, ocular pemphigoid, Lyell's syndrome, neovascularization induced by viral, bacterial, fungal, or parasitic infections, contact lens-induced neovascularization, ulcers, alkali burns, stem cell deficiency, pinguecula, neovascular glaucoma, dry eye disease, Sjogren's syndrome, meibomian gland dysfunction, Stevens-Johnson syndrome, and ocular tumors.

The terms "treatment," "treating," "treat," and the like are used herein to generally refer to obtaining a desired pharmacological and/or physiological effect. This effect may be prophylactic, in that it is a complete or partial prevention of the disease or its symptoms, and/or it may be therapeutic, in that it is a partial or complete stabilization or cure of the disease and/or side effects caused by the disease. The term "treatment" encompasses any treatment of disease in a mammal, particularly a human, and includes (a) preventing the disease and/or symptoms from developing in a subject who may be predisposed to developing the disease or symptoms but has not yet been diagnosed as having the disease or symptoms; (b) inhibiting the disease and/or symptoms, i.e., arresting their progression; or (c) alleviating the disease symptoms, i.e., causing the disease and/or symptoms to regress. Those in need of treatment include those who already have the disease (e.g., those with increased corneal neovascularization, etc.) and those in whom prevention is desired.

Nintedanib {methyl(3Z)-3-{[(4-{methyl[(4-methylpiperazin-1-yl)acetyl]amino}phenyl)amino](phenyl)methylidene}-2-oxo-2,3-dihydro-1H-indole-6-carboxylate} is a kinase inhibitor as described herein. Nintedanib primarily inhibits receptor tyrosine kinases, including, for example, vascular endothelial growth factor receptors (VEGFR1-3), platelet-derived growth factor receptors (PDGFRα and β), and fibroblast growth factor receptors (FGFR1-4).

Formulations and Dosage Regimens The methods described herein include the manufacture and use of pharmaceutical compositions that contain a compound identified by the methods described herein as an active ingredient. Pharmaceutical compositions themselves are also included.

Pharmaceutical compositions typically include a pharma- ceutically acceptable excipient. As used herein, the term "pharma- ceutically acceptable excipient" or "pharma- ceutically acceptable carrier" includes saline, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, suitable for pharmaceutical administration.

As used herein, the phrase "pharmaceutically acceptable salt" means a salt of a compound of interest that is safe and effective for administration to a mammal and possesses the desired biological activity. Pharmaceutically acceptable acid salts include, but are not limited to, hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, superphosphate, I 0 isonicotinate, carbonate, bicarbonate, acetate, lactate, salicylate, citrate, tartrate, propionate, butyrate, pyruvate, oxalate, malonate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., I,I' methylene-bis-(2-hydroxy-3-naphthoate)) salts. Suitable base salts include, but are not limited to, 15 aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, bismuth, and diethanolamine salts.

Methods for formulating suitable pharmaceutical compositions are known in the art, see, for example, Remington: The Science and Practice of Pharmacy, 21st ed., 2005 and Drugs and the Pharmaceutical Sciences: a Series of Textbooks and Monographs (Dekker, NY). For example, solutions, suspensions, creams, ointments, gels, gel-forming solutions, suspensions containing liposomes or micelles, spray formulations, or emulsions used in ophthalmic applications may contain the following components: a sterile diluent, such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol, or other synthetic solvents; antibacterial agents; antioxidants; chelating agents; buffers, such as acetic acid, citric acid, or phosphate, and isotonicity adjusters, such as sodium chloride or dextrose. The pH may be adjusted using an acid or base, such as hydrochloric acid or sodium hydroxide.

The pharmaceutical compositions disclosed herein may contain a "therapeutically effective amount" of an agent described herein. Such an effective amount may be determined based on the effect of the agent administered, or the combined effect of the agents if two or more agents are used. A therapeutically effective amount of an agent may also vary depending on factors such as the disease state, age, sex, and weight of the individual, and the ability of the compound to induce a desired response in the individual, such as improvement of at least one disorder parameter or improvement of at least one symptom of the disorder. A therapeutically effective amount is also an amount in which any toxic or adverse effects of the composition are outweighed by its therapeutically beneficial effects.

Effective doses of the compositions of the present disclosure for the treatment of a condition will vary depending on many different factors, including the means of administration, the target site, the physiological condition of the subject, whether human or animal, other medications being administered, and whether the treatment is prophylactic or therapeutic. Treatment doses may be titrated using conventional methods known to those of skill in the art to optimize safety and efficacy.

Pharmaceutical compositions suitable for injectable use may include sterile aqueous solutions (if water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. It should be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier may be a solvent or dispersion medium, including, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. Proper fluidity may be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. Prevention of the action of microorganisms may be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many instances, it is preferable to include isotonic agents, for example, sugars, polyalcohols, for example, mannitol, sorbitol, and sodium chloride, in the composition. Prolonged absorption of the injectable composition may be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by adding the required amount of the active compound in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by sterile filtration. In general, dispersions are prepared by adding the active compound to a sterile vehicle containing a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying, which yield a powder of the active ingredient and any additional desired ingredients from a previously sterile-filtered solution thereof.

In one embodiment, the therapeutic compound is prepared with a carrier that will protect the therapeutic compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Such formulations can be prepared using standard techniques and are commercially available.

The pharmaceutical compositions may be contained in a container, pack, or dispenser together with instructions for administration.

Nintedanib compositions and formulations may be administered topically or by insertion of a semi-solid formulation or solid implant, or by any other suitable method known in the art. While the agents disclosed herein can be used therapeutically as such, it may be preferable to administer the agents as a pharmaceutical formulation, e.g., mixed with appropriate pharmaceutical excipients, diluents, or carriers selected with respect to the intended route of administration and standard pharmaceutical practice. Pharmaceutical formulations include at least one active compound together with pharma- ceutical acceptable excipients, diluents, and/or carriers.

Administration of the composition or formulation may be once a day, twice a day, three times a day, four times a day, or more. The frequency may be reduced during the maintenance phase of treatment, e.g., once every two or three days instead of every day or twice a day. The dosage and frequency of administration may be adjusted based on the judgment of the treating physician, e.g., taking into account the clinical signs, pathological signs, and clinical and subclinical symptoms of the disease of the condition being treated by the methods of the invention, and the patient's medical history.

It will be understood that the amount of the agents disclosed herein required for use in treatment will vary with the route of administration, the nature of the condition requiring treatment, and the age, weight, and condition of the patient, and is ultimately at the discretion of the attending physician. The compositions typically contain an effective amount of nintedanib. Preliminary doses can be determined according to animal tests, and scaling of doses for administration to humans can be performed according to art-accepted practices.

The length of treatment, i.e., number of days, is readily determined by the physician treating the subject, but the number of days of treatment can range from about 1 day to about 365 days. As provided by the methods of the present invention, the effectiveness of the treatment can be monitored during the course of treatment to determine whether the treatment is successful or whether additional (or modified) treatment is required.

The dosage, toxicity, and therapeutic efficacy of therapeutic compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, to determine the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). Nintedanib dosage forms can be readily determined by one of skill in the art, for example, and can be obtained in animal models and clinical studies reported in the literature for determining dosage, safety, and efficacy according to standard methods known in the art. The specific formulation, route of administration, and dosage can be selected by the individual physician in light of the patient's condition.

Compositions for use in the methods of the invention may contain nintedanib at a concentration of 0.001% to 10% of the total weight or volume of the composition. For example, aqueous compositions may contain 0.001%, 0.01%, 0.1%, 0.5%, 1.0%, 1.5%, 2.0%, 5.0%, or up to 10% nintedanib.

As is well known to those of skill in the art, administration of an aqueous solution to the eye can be in the form of an "eye drop" or multiple drops (e.g., of a nintedanib solution) from a dropper or pipette or other dedicated sterile device. Such drops can typically be up to 50 microliters in volume, but can also be less, e.g., less than 10 microliters.

The present invention is further described in the following examples, which are not intended to limit the scope of the invention described in the claims.

Example 1: Rabbit corneal suture model The rabbit corneal suture model of neovascularization demonstrates the ability of the present method to reduce abnormal corneal neovascularization.

Topical Ophthalmic Formulations Topical compositions were prepared containing 0.2% or 0.05% nintedanib in 10% 2-hydroxypropyl beta-cyclodextrin in phosphate buffer solution, pH 7.4.

Animals and treatments
Thirteen female New Zealand White rabbits were used in this study. Briefly, on day 1, five sutures were placed on the upper cornea of the right eye of each animal to induce neovascularization. Both eyes of these animals were treated with either drugs, vehicle, or saline as described in Table 1.

BID：1日2回（およそ10～12時間間隔）。TID：1日3回（およそ6～8時間間隔）。OD=右眼。OS=左眼。
両眼に投薬し、投薬容量はおよそ40μL/眼であった。
注：第1日の生理食塩水の1回目の投薬は、縫合糸の設置から4時間後に行った。 (Table 1)

BID: twice daily (approximately 10-12 hours apart). TID: three times daily (approximately 6-8 hours apart). OD=right eye. OS=left eye.
Both eyes were dosed and the dose volume was approximately 40 μL/eye.
Note: The first saline dose on day 1 was administered 4 hours after suture placement.

During the study, animals were closely observed for various ocular indications and general physical condition, including body weight. Eye images were taken on days 7, 10, 12, 14, 21, and 28 for analysis of hyperemia.

Data analysis
The eye images were analyzed using NIH ImageJ® software. Each image was opened in ImageJ®, the ruler was used to mark the photograph, and the neovascularized area on the cornea near the suture was selected with the selection tool. The area in ^mm2 was calculated with the measurement tool in the software and recorded in Excel, and the image was captured and saved. A two-tailed T-test was used to determine whether there were significant differences between groups. The results were plotted as a histogram of the mean with standard deviation for ease of comparison.

Results and Discussion As shown in Figures 2A and 2B, nintedanib showed a significant inhibitory effect on suture-induced neovascularization in rabbit corneas 12 and 14 days after suture induction. A higher dose of 0.2% nintedanib showed better efficacy than 0.05% nintedanib, and a more frequent dosing regimen of TID dosing showed greater efficacy compared to BID dosing.

In summary, nintedanib potently inhibited suture-induced corneal neovascularization. This strong activity is likely due to the special targeting profile of nintedanib, which has potent activity against VEGFR1-3 and FGFR1-2. These results support the methods disclosed herein.

Example 2: Mouse Corneal Graft Rejection Model In this example, C57BL/6 mouse corneas are transplanted onto BALB/c mouse corneas as described (Sonoda et al. Invest Ophthalmol Vis Sci. 1995 Feb; 36(2): 427-34; Invest Ophthalmol Vis Sci. 2000 Mar; 41(3): 790-8; Yamagami et al. Invest Ophthalmol Vis Sci. 2001 May; 42(6): 1293-8). Seven days after transplantation, the sutures are removed. Mice are divided into two groups: group 1 is treated with nintedanib 0.2% solution and group 2 is treated with vehicle solution. Treatment begins immediately after transplantation and is performed TID for 8 weeks. Corneal opacity and graft rejection are evaluated weekly for 8 weeks as described.

The nintedanib-treated group showed significantly higher graft survival (lower rejection rate) over the 8-week study. This result indicates that nintedanib 0.2% solution can prevent corneal graft rejection.

Example 3: Formulation Nintedanib Ophthalmic Solution This drug formulation is an isotonic ophthalmic solution prepared with 2-hydroxypropyl beta-cyclodextrin or other similar cyclodextrin and a buffer solution with a pH range of 5.5 to 8.0. Other viscosity agents, lubricants, preservatives may be added to enhance the functionality of the formulation. The composition of this ophthalmic solution is shown in Table 2.

Table 2. Nintedanib ophthalmic solution

Nintedanib Ophthalmic Suspension This drug formulation is an isotonic ophthalmic suspension prepared with sodium carboxymethylcellulose and a buffer solution with a pH range of 5.5 to 8.0. The drug particle size is less than 40 micrometers. Other viscosity agents, lubricants, solubilizers, and preservatives may be added to enhance the functionality of this formulation suspension. Its composition is shown in Table 3.

Table 3. Nintedanib Ophthalmic Suspension

Nintedanib Ophthalmic Emulsion This drug formulation is an isotonic ophthalmic emulsion. The drug is dissolved in a mixed oil phase and emulsifying excipients, which are then emulsified and mixed with an aqueous phase with a pH range of 5.5 to 8.0. Other viscosity agents, lubricants, solubilizers, and preservatives may be added to enhance the functionality of this emulsion formulation. Its composition is shown in Table 4.

Table 4. Nintedanib Ophthalmic Emulsion

Nintedanib sustained release semi-solid formulation This drug formulation is an isotonic sustained release semi-solid formulation. The drug is dissolved and/or suspended in a semi-solid medium with a pH range of 5.5 to 8.0. Other viscosity agents, lubricants, solubilizers, and preservatives may be added to enhance the functionality of this sustained release semi-solid formulation. Its composition is shown in Table 5.

Table 5. Sustained release semi-solid formulations

Nintedanib sustained release implant This drug formulation is a solid implant. The drug is mixed and blended with one or more polymers. The mixture of drug and polymer is melted at a predetermined temperature and extruded into a filament with a predetermined diameter size. The filament formulation is cut into a segment of a predetermined size that can be implanted into ocular tissue. Its composition is shown in Table 6.

Table 6. Sustained release implants

Without limitation, the example compositions used in the methods according to the present invention may be modified from the ophthalmically acceptable compositions shown.

Other Aspects Although the present invention has been described with reference to its detailed description, it should be understood that the above description is intended to be illustrative and not limiting of the scope of the invention as defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the appended claims.

Claims (8)

1. A composition for treating an ophthalmic indication involving abnormal neovascularization in the anterior segment of the eye of a subject, comprising an effective amount of nintedanib or a pharma- ceutical acceptable salt thereof,
the ophthalmic indication being graft-versus-host disease; atopic conjunctivitis; ocular rosacea; ocular pemphigoid; Lyell's syndrome; neovascularization induced by viral, bacterial, fungal, or parasitic infection; contact lens-induced neovascularization; ulcers; alkali burns; or stem cell deficiency ;
The composition, wherein the composition is in the form of a topical ophthalmic formulation or in the form of an ocular implant. 13. The composition of claim 1 in the form of an ocular implant. The composition of claim 2, wherein the ocular implant is in the form of a semi-solid or solid sustained release implant. The composition of claim 2, wherein the ocular implant is inserted into the eye of the subject. The composition of claim 1, wherein the topical ophthalmic formulation is a solution, suspension, cream, ointment, gel, gel-forming liquid, suspension containing liposomes or micelles, spray formulation, or emulsion. The composition of claim 1, wherein the topical ophthalmic preparation is a topical eye drop. 10. The composition of claim 1, in the form of a topical ophthalmic formulation, wherein the topical ophthalmic formulation is a topical eye drop. The composition of claim 7 , wherein the composition is administered locally to the eye of the subject .

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